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Researchers who aim to globally analyze the gastrointestinal immune system via flow cytometry have many protocol options to choose from, with specifics generally tied to gut wall layers of interest. To get a clearer idea of the approach we should use on full-thickness colon samples from mice, we first undertook a systematic comparison of three tissue dissociation techniques: two based on enzymatic cocktails and the other one based on manual crushing. Using flow cytometry panels of general markers of lymphoid and myeloid cells, we found that the presence of cell-surface markers and relative cell population frequencies were more stable with the mechanical method. Both enzymatic approaches were associated with a marked decrease of several cell-surface markers. Using mechanical dissociation, we then developed two minimally overlapping panels, consisting of a total of 26 antibodies, for serial profiling of lymphoid and myeloid lineages from the mouse colon in greater detail. Here, we highlight how we accurately delineate these populations by manual gating, as well as the reproducibility of our panels on mouse spleen and whole blood. As a proof-of-principle of the usefulness of our general approach, we also report segment- and life stage-specific patterns of immune cell profiles in the colon. Overall, our data indicate that mechanical dissociation is more suitable and efficient than enzymatic methods for recovering immune cells from all colon layers at once. Additionally, our panels will provide researchers with a relatively simple tool for detailed immune cell profiling in the murine gastrointestinal tract, regardless of life stage or experimental conditions.

We aim to investigate the correlations between hemodynamic parameters, penile rigidity grading, and the therapeutic effects of phosphodiesterase type 5 inhibitors using color Doppler flow imaging after intracavernosal injection in patients with erectile dysfunction. This study involved 164 patients. After intracavernosal injection with a mixture of papaverine （60 mg）, prostaglandin E1 （10 μg）, and lidocaine （2%, 0.5-1 ml）, the penile vessels were assessed using color Doppler flow imaging. Penile rigidity was classified based on the Erection Hardness Score system as Grades 4, 3, 2 or 1 （corresponding to Schramek Grades Ⅴ to Ⅱ）. Then, the patients were given oral sildenafil （50-100 mg） and scored according to the International Index of Erectile Function （IIEF-5） questionnaire. The number of patients with penile rigidities of Schramek Grades Ⅱ to Ⅴ was 14, 18, 21, and 111, respectively. The IIEF-5 score was positively correlated with the refilling index of the penile cavernosal artery （r = 0.79, P 〈 0.05）, the peak systolic velocity （r= 0.45, P〈 0.05）, and penile rigidity （r= 0.75, P〈 0.05）, and was negatively correlated with the end diastolic velocity （r = -0.74, P 〈 0.05）. For patients with erectile dysfunction, both the IIEF-5 score after sildenafil administration, which is correlated with penile rigidity, and the hemodynamic parameters detected using color Doppler flow imaging may predict the effects of phosphodiesterase type 5 inhibitor treatment and could provide a reasonable model for the targeted-treatment of erectile dysfunction.

This paper describes and tests a method for automatically determining the envelope of the blood flow velocity curve in ultrasonic Doppler imaging, which was implemented in a prototype of the 128-channel mobile ultrasound B-mode scanner. On the basis of the determined envelopes, algorithms were also developed for the automatic determination of the most important characteristic points of the Doppler blood flow spectrum in pulse wave Doppler imaging mode and the most relevant blood flow parameters. Sufficiently good repeatability and precision were obtained with low computational complexity.

Immune inflammatory response plays an important role in chronic obstructive pulmonary disease (COPD). However, the cellular immune status of patients with COPD at different phases is unclear. Herein, we aim to investigate the distribution and functional status of T cell subsets in different phases of COPD (acute exacerbation of COPD [AECOPD] and stable COPD [SCOPD]). This is an observational case-control study undertaken in West China Hospital. The distribution of T cell subsets in peripheral blood of AECOPD, SCOPD, and healthy controls (HCs) was measured using multi-color flow cytometry, and the functional status was analyzed by additional staining of activation markers. A total of 43 HCs, 43 SCOPD patients, and 64 AECOPD patients were evaluated. The total number and percentage of lymphocytes and the CD4+/CD8+ T cells ratio were significantly lower in AECOPD patients when compared to HCs. HLA-DR expression in CD3+, CD4+, CD8+, CD8+ TCR aβ, and CD4+ TCR aβ T cells was upregulated in the AECOPD group. Similarly, the expressions of HLA-DR, CD57, and PD-1 were higher in T cell subsets in the AECOPD group. Compared with the SCOPD and HC groups, the AECOPD had a significantly lower proportion of CD4+CD27+CD28+ T cells, but opposite results were found for CD4+CD27-CD28- T cells. In addition, the proportion of CD4+CD39+ T cells and CD4+CD25+FoxP3+ T cells was significantly higher in the AECOPD and SCOPD groups when compared to the HC group ( < 0.05). The distribution of nearly half the T cell subsets in AECOPD patients was significantly different from that in SCOPD patients and HCs. AECOPD patients may have cellular immune suppression, immune dysfunction, abnormal activation, and higher senescence depletion of T cells.

Background： Endovascular aneurysnl repair （EVAR） is one of the first-line therapies of abdominal aortic aneurysms. Postoperative endoleak is the most common complication of EVAR. Computed tomography angiography （CTA）, which is routine for follow-up, has side effects （e.g., radiation） and also has a certain percentage of missed diagnosis. Preliminary studies on contrast-enhanced ultrasound （CEUS） have shown that the sensitivity of CEUS for detecting endoleak is no lower than that of CTA. To investigate the advantages of CEUS, we conducted CEUS examinations of post-EVAR cases in which CTA failed to detect endoleak or could not verify the type of endoleak. Methods： Post-EVAR patients, who were clinically considered to have endoleak and met the inclusion criteria were enrolled between March 2013 and November 2014. All of the patients underwent color Doppler flow imaging （CDFI） and a CEUS examination. Size, location, microbubble dispersion, and hemodynamic characteristics of leaks were recorded. Comparison between the diagnosis of CEUS and CDF1 was conducted using Fisher＇s exact test and clinical outcomes of all patients were followed up. Results： Sixteen patients were enrolled, and 12 （75%） had endoleaks with verified types by CEUS. Among 12 cases of endoleaks were positive by CEUS, 10 were CDFl-positive, and the four CEUS-negative cases were all negative by CDFI. The diagnostic values of CEUS and CDFI were statistically difl＇erent （P = 0,008）. Six patients with high-pressure endoleaks received endovascular re-intervention guided by CEUS results. One patient with type Ⅲ endoleak had open surgery when endovascular repair failed. Conclusions： C EUS is a new, safe, and effective means for detection ofendoleaks post-EVAR. This technique can be used as a supplement for routine CTA follow-up to provide more detailed information on endoleak and its category.

Intubation is a frequently performed procedure in emergency medicine that is associated with significant morbidity and mortality when unrecognized esophageal intubation occurs. However, it may be difficult to visualize the endotracheal tube (ETT) in some patients. This study assessed whether the addition of color Doppler was able to improve the ability to visualize the ETT location. This study was performed in a cadaver lab using three different cadavers chosen to represent varying neck circumference. Cadavers were randomized to tracheal or esophageal intubation. Blinded sonographers then assessed the location of the ETT using either grayscale or color Doppler imaging. Accuracy of sonographer identification of ETT location, time to identification, and operator confidence were assessed. One hundred and fifty intubations were performed and each was assessed by both standard and color Doppler techniques. There were 78 tracheal intubations and 72 esophageal intubations. The standard technique was 99.3% (95% CI 96.3 to 99.9%) accurate. The color flow technique was also 99.3% (95% CI 96.3 to 99.9%) accurate. The mean operator time to identification was 3.24s (95% CI 2.97 to 3.51s) in the standard approach and 5.75s (95% CI 5.16 to 6.33s) in the color flow technique. The mean operator confidence was 4.99/5.00 (95% CI 4.98 to 5.00) in the standard approach and 4.94/5.00 (95% CI 4.90 to 4.98) in the color flow technique. When added to standard ultrasound imaging, color flow did not improve accuracy or operator confidence for identifying ETT location and resulted in a longer examination time.

Objective The ultrasonic diagnosis of cervical and facial cystic masses, as well as cases of missed diagnosis and misdiagnosis, was examined, to improve the diagnosis of branchial cleft anomalies. Methods A retrospective analysis was conducted on 17 patients with branchial cleft cyst anomalies, including 11 males and 6 females, aged 12–53 years, with an average age of 33 ± 2 years, were unilateral single. All patients who underwent an ultrasound examination and image storage for retrospective analysis, and both longitudinal and transverse sections were scanned to observe the shape, size, boundary, peripheral relationship, and blood flow signal of the masses. All cases were examined with an enhanced CT scan, and pathological reports were generated. Results Among the 17 cases of branchial cleft anomalies, 15 cases were branchial cleft cysts, while one case involved fistula formation and one case involved sinus tract formation. Based on the type of branchial cleft, the first, second, and third cysts were classified in 4, 12, and 1 case, respectively. The sensitivity rate and specificity of ultrasonic diagnosis were 14/17 (82.4%) and 4/6 (66.7%), respectively. Ultrasonic characteristic analysis for the masses can be found in simple cystic masses or hypoechoic masses, most of them are of a regular shape and have a distinct boundary, and almost no blood flow signal. All patients who were misdiagnosed exhibited blood flow signals, including 1 patient with an abundant blood flow signal, 1 patient suspected of having ectopic thyroid with an abnormal function due to the rat-tail sign, 2 patients misdiagnosed as local inflammatory focus, and 1 patient misdiagnosed with tuberculous lymphadenitis. Conclusion Ultrasound has a detection rate of up to 100% for cervical and facial masses, providing a fundamental determination of lesion characteristics and specific guidance for preoperative diagnosis. If the blood flow signals can be identified and carefully considered their peripheral relationship, the diagnostic rate can be improved.

Background: Thyrotoxicosis is a metabolic state of excessive thyroid hormones which include tetraiodothyronine (FT4) and/or triiodothyronine (FT3). The most common causes of thyrotoxicosis are either autoantibody-mediated hyperfunction of thyroid gland (Graves’ disease) or inflammation-induced destructive thyroiditis (subacute thyroiditis and postpartum thyroiditis). The correct diagnosis of thyrotoxicosis is important as management differs according to etiology. The radioactive iodine uptake test and technetium pertechnetate thyroid scan are the gold standard for diagnosis of thyrotoxicosis. However, these tests are limited by their availability and high cost. Therefore, it is important to look for other parameters which can help in the diagnosis of hyperthyroidism. The non-invasive thyroid color flow Doppler sonography (CFDS) is an easily available, highly informative, safe, and comparatively low-cost investigation which can be used as an alternative. Aims and Objectives: The aim of the study was to assess the role of CFDS in the differential diagnosis of thyrotoxicosis. Materials and Methods: A total number of 120 newly diagnosed thyrotoxic patients were enrolled. On the basis of clinical, laboratory evaluation, and thyroid scan, patients were diagnosed with either Graves’ disease or thyroiditis. All the patients underwent CFDS in the radiology department. Results: Ninety patients of Graves’ disease and thirty patients of thyroiditis were enrolled in our study. The mean peak systolic velocity (PSV) of inferior thyroid artery (ITA) from both lobes of thyroid gland were used as markers of thyroid vascularity. The difference in mean PSV-ITA in Graves’ and thyroiditis patients was statistically significant (P < 0.001). Using receiver operating characteristic analysis for diagnostic accuracy of PSV, we demonstrated that the cutoff 50 cm/s had 98.9% sensitivity and 96.7% specificity in differentiating Graves’ disease from thyroiditis with positive predictive value and negative predictive value being 99% and 96%, respectively. Conclusion: A non-invasive CFDS is an inexpensive and easily available procedure which provides the functional status of the thyroid gland. The PSV of ITA can help to differentiate the etiologies of thyrotoxicosis and can be considered as an alternative to nuclear scanning where such modality is not present or contraindicated.

Vessel diseases are often accompanied by abnormalities related to vascular shape and size. Therefore, a clear visualization of vasculature is of high clinical significance. Ultrasound color flow imaging (CFI) is one of the prominent techniques for flow visualization. However, clutter signals originating from slow-moving tissue are one of the main obstacles to obtain a clear view of the vascular network. Enhancement of the vasculature by suppressing the clutters is a significant and irreplaceable step for many applications of ultrasound CFI. Currently, this task is often performed by singular value decomposition (SVD) of the data matrix. This approach exhibits two well-known limitations. First, the performance of SVD is sensitive to the proper manual selection of the ranks corresponding to clutter and blood subspaces. Second, SVD is prone to failure in the presence of large random noise in the dataset. A potential solution to these issues is using decomposition into low-rank and sparse matrices (DLSM) framework. SVD is one of the algorithms for solving the minimization problem under the DLSM framework. Many other algorithms under DLSM avoid full SVD and use approximated SVD or SVD-free ideas which may have better performance with higher robustness and less computing time. In practice, these models separate blood from clutter based on the assumption that steady clutter represents a low-rank structure and that the moving blood component is sparse. In this paper, we present a comprehensive review of ultrasound clutter suppression techniques and exploit the feasibility of low-rank and sparse decomposition schemes in ultrasound clutter suppression. We conduct this review study by adapting 106 DLSM algorithms and validating them against simulation, phantom, and in vivo rat datasets. Two conventional quality metrics, signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR), are used for performance evaluation. In addition, computation times required by different algorithms for generating clutter suppressed images are reported. Our extensive analysis shows that the DLSM framework can be successfully applied to ultrasound clutter suppression.

Efficient imaging of blood flow disturbances resulted from carotid atherosclerosis plays a vital role clinically to predict brain stroke risk. Carotid atherosclerosis and its development is closely linked with raised blood viscosity. Therefore, study of viscosity changing hemodynamic effect has importance and it might be useful for improved examination of carotid atherosclerosis incorporating the viscosity induced contrast in conventional ultrasound imaging. This work considered the design of realistic models of atherosclerotic carotid artery of different stages and solved to compute the hemodisturbances using computational fluid dynamics (CFD) by finite element method (FEM) to investigate viscosity changes effect. Ultrasound color flow image of velocities of blood have been constructed using phase shift information estimated with autocorrelation of Hilbert transformed simulated backscattered radiofrequency (RF) signals from moving blood particles. The simulated ultrasound images have been compared with CFD simulation images and identified a good match between them. The atherosclerosis stages of the models have been investigated from the estimated velocity data. It has been observed that the blood velocities increase noticeably in carotid atherosclerotic growths and velocity distribution changes with viscosity variations. It is also found importantly that the viscosity induced contrast associated to atherosclerosis is detectable in ultrasound color flow imaging. The findings of this work might be useful for better investigation of carotid atherosclerosis as well as prediction of its progression to reduce the stroke risk.